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  • Immunohematology


Article | 16-October-2019

Low-ionic-strength saline solution–antiglobulin test (LISS-AGT)

The use of low-ionic-strength saline (LISS) solution as an enhancement for antibody screening and crossmatching was first described by Löw and Messeter in 1974. This method allowed for a reduced incubation time while maintaining adequate specificity and sensitivity of the antiglobulin test (AGT). Since then, the LISS-AGT tube method has been widely used in antibody detection and identification, as well as compatibility testing. As initially described, the method used red blood cells

LeeAnn Walker

Immunohematology, Volume 34 , ISSUE 2, 57–60

Article | 14-October-2020

The investigation of the significance of a positive direct antiglobulin test in blood donors

samples were not included in this study. With a tube test, most of the antibodies had titers from 4 to 8. IgG subclass studies showed that 14 of 25 samples with reactive eluates contained IgG1, one contained IgG1+IgG2, one contained IgG1+IgG4, and two contained IgG1+IgG3 weak. The frequency of donors with a positive direct antiglobulin test (DAT) was ~ 1 in 3000 and males were twice as likely to be DAT positive (8 females vs. 17 males in this study). None of the donors had hemolysis. Two donors showed

Marianna Bellia, John Georgopoulos, Vasilis Tsevrenis, Efrosini Nomikou, Niki Vgontza, I. Kontogpoulous-Griva

Immunohematology, Volume 18 , ISSUE 3, 78–81

Article | 14-October-2020

Clinically significant autoimmune hemolytic anemia with a negative direct antiglobulin test by routine tube test and positive by column agglutination method

In two to five percent of cases of autoimmune hemolytic anemia (AIHA),the patient’s RBCs are negative in the direct antiglobulin test (DAT). We describe a patient with AIHA with a negative DAT when tested by the traditional tube test but with a strongly positive one when tested by column agglutination. When the DAT was repeated by tube test using 4°C saline washes, it became positive. This phenomenon has been observed when low-affinity antibodies are involved in AIHA. The patient

Marco Lai, Carlo Rumi, Giuseppe D’Onofrio, Maria Teresa Voso, Giuseppe Leone

Immunohematology, Volume 18 , ISSUE 4, 109–113

Article | 18-October-2020

Frequency of HLA-DQB*06 in Caucasian, African American, and Mexican American patients with a positive direct antiglobulin test

A reduced frequency of HLA-DQ6 in patients with a positive direct antiglobulin test (DAT) was previously reported but race was undisclosed. Therefore, we investigated a total of 275 patients (80 Caucasian, 113 African American, and 82 Mexican American) and 518 normal controls (205 Caucasian, 208 African American, and 105 Mexican American). These were typed for class II HLA antigens using molecular techniques. A DAT was performed on each patient’s red cells drawn into EDTA using both mouse

Joann M. Moulds, Laura A. Diekman, T. Denise Wells, John D. Reveille

Immunohematology, Volume 16 , ISSUE 2, 74–77

Article | 15-February-2021

Albumin-indirect antiglobulin test

reagent RBC suspension. Centrifuge, and perform immediate spin reading (if desired). Add two drops albumin (per MI). Incubate 30–60 minutes at 37°C (per MI). Centrifuge, and perform 37°C reading. Wash three to four times in isotonic saline. Add one to two drops AHG reagent (per MI). Centrifuge, and perform AHG reading. Compiled from Fung et al.6 RBC = red blood cell; MI = manufacturer’s instructions; AHG = antihuman globulin. Indications The albumin-indirect antiglobulin test (IAT) is a

J.R. Hamilton

Immunohematology, Volume 35 , ISSUE 2, 63–64

Article | 16-February-2021

Saline–indirect antiglobulin test

Principle The saline–indirect antiglobulin test (IAT) or saline– indirect Coombs test refers to a test incubated at 37°C without enhancement media before washing and the addition of antihuman globulin (AHG) reagent. Saline-IAT tests rely solely on the binding kinetics between plasma antibodies and antigens expressed on red blood cells (RBCs) suspended in isotonic saline. In this environment, the Na+ in the saline may partially neutralize the oppositely charged RBC membrane, blocking the

J.R. Hamilton

Immunohematology, Volume 35 , ISSUE 4, 156–158

Review | 01-December-2019

EDTA glycine acid treatment of red blood cells

IgG dissociation is necessary when a sample is direct antiglobulin test (DAT) positive and antigen testing using blood grouping serum reactive by the antiglobulin test is performed. Exposure of IgG-coated red blood cells (RBCs) to a low pH of 3.0 with EDTA glycine acid successfully dissociates the IgG, rendering the RBCs DAT negative 82 to 85 percent of the time. The procedure takes one minute or less and leaves RBC antigens intact and able to be typed except for those antigens in the Kell

Joanne Kosanke

Immunohematology, Volume 28 , ISSUE 3, 95–96

Article | 13-April-2020

Problems highlighted when using anticoagulated samples in the standard tube low ionic strength antiglobulin test

Amanda J Sweeney

Immunohematology, Volume 22 , ISSUE 2, 72–77

Article | 16-October-2019

Utility of chloroquine diphosphate in the blood bank laboratory

Chloroquine diphosphate (CDP) is a helpful tool in the blood bank for two main applications. The most common application is to render direct antiglobulin test–positive red blood cells (RBCs) free from membrane-bound IgG; these treated RBCs can then be used for autologous adsorption and/or to determine the patient’s RBC phenotype. Another common use of CDP is to remove human leukocyte antigens (HLAs) from RBCs to help identify or exclude the presence of antibodies to HLAs expressed

Thandar Aye, Patricia A. Arndt

Immunohematology, Volume 34 , ISSUE 3, 98–102

Article | 22-November-2020

Use of monoclonal Jka and Jkb reagents in phenotyping red cells with a positive direct antiglobulin test

Twenty-five red cell samples with a positive anti-IgG direct antiglobulin test QAT) were tested with human monoclonal IgM Jka and Jkb antibodies. Red cell samples were first tested by a 5- minute incubation tube test with the monoclonal antibodies (MAbs). The same red cells were then chloroquine diphosphate (CDP)-treated, and retested. Eleven of the CDP-treated samples were also tested with conventional polyclonal antibodies (PAhs) that required a 37°C incubation for 30 minutes, followed by

Judy L. Brazell

Immunohematology, Volume 10 , ISSUE 1, 16–18

Article | 20-December-2020

Du phenotyping by the rosette technique when the direct antiglobulin test is positive

The rosette technique provides a simple rapid and accurate procedure for Du phenotyping of red blood cells (RBCs) with a positive direct antiglobulin test (DAT) due to antibodies other than those of the Rh system. We used Sebring and Polesky’s rosette technique to test RBCs with positive DATs. Characteristic microscopic agglutination was observed for the Du phenotype which was dif­ferent from the mixed-field agglutination seen as positive rosettes in feto-maternal hemorrhage (FMH

Jochewed Werch, Carolyn Todd, Marilyn K. Moulds

Immunohematology, Volume 6 , ISSUE 2, 44–46

Article | 09-November-2020

A maternal warm-reactive autoantibody presenting as a positive direct antiglobulin test in a neonate

Autoimmune hemolytic anemia in pregnancy is a rare cause of hemolytic disease of the newborn. This report describes a neonate with a mild hemolytic process and a positive direct antiglobulin test (DAT) presenting as the first manifestations of a maternal warm-reactive autoantibody. A full-term male neonate, blood group O, had a strongly positive DAT and laboratory evidence suggestive of a mild hemolytic process. The neonate’s mother was also group O and had a negative antibody screen

Terry D. Williamson, Linda H. Liles, Douglas P. Blackall

Immunohematology, Volume 13 , ISSUE 1, 6–8

Case report | 14-October-2020

Autoanti-D in a patient after cladribine treatment for lymphoplasmocytic lymphoma

We report the case of a 62-year-old woman who developed an autoanti-D after cladribine treatment. In May 2000, the patient underwent splenectomy for a stage IV-B lymphoplasmocytic lymphoma. She was transfused with ABO- and Rh(D)-matched blood. A month later, she received chemotherapy with cladribine. In February 2001, blood grouping showed her to be AB, D+ and the direct antiglobulin test was positive for IgG. An autoanti-D was identified in the eluate. Genotypic analysis confirmed the Rh

Joan Cid, Victor Beltran, L. Escoda, Enric Elies, Carmen Martin-Vega

Immunohematology, Volume 18 , ISSUE 1, 16–18

Article | 26-October-2020

A comparison of a new affinity column system with a conventional tube LISS-antiglobulin test for antibody detection

A recently introduced system for antibody detection (ReACT™) consists of affinity columns (AFC) that contain protein A and protein Gcoated agarose. We compared the ReACT™ system to a conventional tube low-ionic-strength saline antiglobulin test (LISS-AGT). We selected 100 LISS-AGT positive samples with clinically important and benign antibodies of varying strengths and 130 LISS-AGT negative samples to evaluate by the AFC method. AFC tests were positive with all 84 clinically

R. Sue Shirey, Joan S. Boyd, Christine Barrasso, Karen E. King, Paul M. Ness

Immunohematology, Volume 15 , ISSUE 2, 75–77


Demonstration of IgG subclass (IgG1 and IgG3) in patients with positive direct antiglobulin tests

Serologic characterization of autoantibodies helps in the management and monitoring of the course of autoimmune hemolytic anemia (AIHA). The purpose of this study was to evaluate gel centrifugation test (GCT) cards for immunoglobulin G (IgG) titer and determination of IgG subclasses IgG1 and IgG3 and their influence on hemolysis. Eighty direct antiglobulin test (DAT)-positive patients were examined with the help of GCT cards for IgG titer and IgG subclasses. The results were correlated with the

Ashutosh Singh, Archana Solanki, Rajendra Chaudhary

Immunohematology, Volume 30 , ISSUE 1, 24–27

Article | 09-November-2020

Measurement of red blood cell-bound C3b and C3d using an enzyme-linked direct antiglobulin test

Complement has a complex role in immune mediated red blood cell (RBC) destruction and usually induces extravascular hemolysis of C3bcoated RBCs by erythrophagocytosis and by acting synergistically with cell-bound immunoglobulins. A sensitive two-stage enzyme-linked direct antiglobulin test (ELDAT) was developed and used to measure RBC-bound C3b and C3d in 120 healthy adult individuals and in 60 patients suffering from a variety of conditions, including warm- and cold-type autoimmune hemolytic

J.D. Bellamy, D.J. Booker, N.T. James, R. Stamps, R.J. Sokol

Immunohematology, Volume 13 , ISSUE 4, 123–131

Report | 09-October-2019

A detailed flow cytometric method for detection of low-level in vivo red blood  cell–bound IgG, IgA, and IgM

when tested with Ig-coated RBCs and the least amount of fluorescence when tested with naive RBCs. Tannic acid was used to prepare Ig-coated RBCs. Cross-reactivity of FITC-conjugated anti-IgG, -IgA, and -IgM with Ig-coated RBCs was evaluated, and a reference range was established. Use of this method may assist in clinical evaluation of patients who present with hemolysis and a negative direct antiglobulin test.

Wendy Beres, Geralyn M. Meny, Sandra Nance

Immunohematology, Volume 32 , ISSUE 4, 161–169

Report | 14-March-2020

Attempts to support an immune etiology in 800 patients with direct antiglobulin test–negative hemolytic anemia

Clinical and hematologic evidence of warm autoimmune hemolytic anemia (AIHA) is present in some patients whose direct antiglobulin test (DAT) is negative. The most common causes for AIHA associated with a negative DAT are RBC-bound IgG below the sensitivity threshold of the DAT, RBC-bound IgA and IgM not detectable by routine reagents, and low-affinity IgG that dissociates during the testing process. Samples submitted from 800 patients with hemolytic anemia and a negative DAT were tested by an

Regina M. Leger, Asuncion Co, Penny Hunt, George Garratty

Immunohematology, Volume 26 , ISSUE 4, 156–160


Clinical and reference lab characteristics of patients with suspected direct antiglobulin test (DAT)-negative immune hemolytic anemia

Clinical evidence of warm autoimmune hemolytic anemia is present in 1 percent to 10 percent of patients whose direct antiglobulin test (DAT) is negative. The clinical underpinnings associated with DAT-negative immune hemolysis are poorly understood, and the current study aimed to further define the clinical characteristics associated with this form of anemia. A 19-question survey, requesting clinical information about each patient, was retrospectively mailed to all referring labs that had sent

Matthew S. Karafin, Gregory A. Denomme, Michael Schanen, Jerome L. Gottschall

Immunohematology, Volume 31 , ISSUE 3, 108–115

Article | 26-October-2020

EDTA/glycine-acid versus chloroquine diphosphate treatment for stripping Bg antigens from red blood cells

EDTA/glycine-acid (EGA) has been reported to remove IgG-bound antibodies from red blood cells (RBCs) and to denature Kell system and Era antigens. EGA-treated RBCs were tested in parallel with chloroquine diphosphate (CDP)-treated RBCs to evaluate whether EGA would remove Bg antigens from RBCs as efficiently as CDP. Fifty-seven serum/plasma samples containing known Bg antibodies were tested with untreated Bg+ RBCs, EGA-treated Bg+ RBCs, and CDP-treated Bg+ RBCs by an indirect antiglobulin test

Kayla D. Champagne, Peggy Spruell, Jane Chen, Leslie Voll, Gloria Schlanser

Immunohematology, Volume 15 , ISSUE 2, 66–68

Review | 01-December-2019

Polyethylene glycol antiglobulin test  (PEG-AGT)

Larry Weldy

Immunohematology, Volume 30 , ISSUE 4, 158–160

Article | 14-October-2020

The direct antiglobulin test in a hospital setting

Joan Cid, Xavier Ortín, Víctor Beltran, Lourdes Escoda, Enric Contreras, Enric Elies, Carmen Martín-Vega

Immunohematology, Volume 19 , ISSUE 1, 16–18


Indirect antiglobulin test-crossmatch using low-ionic-strength saline–albumin enhancement medium and reduced incubation time: effectiveness in the detection of most clinically significant antibodies and impact on blood utilization

Indirect antiglobulin test-crossmatch (IAT-XM) using enhancement media such as low-ionic-strength saline (LISS) and polyethylene glycol (PEG) usually requires 15 minutes of incubation. These methods are necessary when testing samples from blood recipients who have a higher risk of alloimmunization. In emergency situations, IAT-XM can be time-consuming and can influence presurgery routine, resulting in more red blood cell (RBC) units being tested and stored to avoid the transfusion of 

Carla Luana Dinardo, Sílvia Leão Bonifácio, Alfredo Mendrone Júnior

Immunohematology, Volume 30 , ISSUE 1, 1–5

Article | 16-October-2019

Dithiothreitol treatment of red blood cells

treatment of multiple myeloma, targets the CD38 antigen on malignant plasma cells. CD38 is also weakly expressed on all normal RBCs, including those in RBC reagents used in pretransfusion testing. This expression complicates the identification of clinically significant RBC antibodies because the plasma/serum of such patients will react with most or all RBCs in antibody detection and identification tests performed by the indirect antiglobulin test.5 Because CD38 configuration depends on disulfide bonds

C.B. Bub

Immunohematology, Volume 33 , ISSUE 4, 170–172

Article | 01-April-2020

Application of gel technology in the serologic characterization of autoantibody in DAT-positive autoimmune diseases

Sudipta Sekhar Das, Rajendra K. Chaudhary

Immunohematology, Volume 23 , ISSUE 2, 59–62

Article | 03-November-2020

Evaluation of column technology for direct antiglobulin testing

Joann M. Moulds, Laura Diekman, T. Denise Wells

Immunohematology, Volume 14 , ISSUE 4, 146–148

Review | 27-December-2020

A review: the enzyme-linked antiglobulin test (ELAT) and its applications for reference laboratories

Ruth Mougey

Immunohematology, Volume 5 , ISSUE 3, 69–78

Article | 15-February-2021

Heat elution: a modification of the Landsteiner-Miller method

eluate and final wash in parallel, using the appropriate test phase(s) in a manner that is best for detection of the antibody, following the laboratory’s standard operating procedure. RBCs = red blood cells. Additionally, heat causes a conformational change in proteins, resulting in a loss of the “lock and key” interaction between antigen and antibody. Indications Elution studies may be performed on patient samples having a positive direct antiglobulin test (DAT) due to auto- or

C. Dean-El, N. Quraishy

Immunohematology, Volume 35 , ISSUE 2, 45–47

Case report

Anti-Jk3 in a Filipino man

ordered for transfusion. During subsequent pre-transfusion compatibility testing, the antibody screen was found to be positive (all screening cells reactive at the antihuman globulin phase). An antibody identification panel was performed. The patient’s serum was found to react with all panel cells tested, including the autocontrol tube. A direct antiglobulin test revealed the presence of both anti-IgG and anti-C3 coating the patient’s RBCs. The specimen was then sent to a reference

Shaina McCaskill, Scott Wise, Sheila Tinsley

Immunohematology, Volume 31 , ISSUE 3, 119–122

Article | 18-October-2020

A quick and simple method for phenotyping IgG-sensitized red blood cells

Positive (IgG) direct antiglobulin test (DAT) reactivity ranging from weakly positive to 2+ can be eliminated using a simple “blocking” technique with anti-IgG. This method can be used for antigen typing DAT-positive red blood cells that require the antiglobulin technique.

Thom S. Sererat, Douglas W. Veidt, Ann M. Dutched

Immunohematology, Volume 16 , ISSUE 4, 154–156

Case report | 01-December-2019

Paroxysmal cold hemoglobinuria: a case report

Scott C. Wise, Sheila H. Tinsley, Lloyd O. Cook

Immunohematology, Volume 28 , ISSUE 4, 118–123

Article | 14-October-2020

Warm autoimmune hemolytic anemia with mimicking anti-c and -E specificities

An 18-month-old male was admitted to a hospital with a hemoglobin of 4.1 g/dL and a reticulocyte count of 53 percent. There was no history of prior transfusion. Serologic evaluation revealed the presence of both a positive direct antiglobulin test (DAT) and an indirect antiglobulin test (IAT). The patient’s red blood cells (RBCs) typed as group A, C–D–E–c+e+ (cde/cde). Evaluation of the IAT revealed the presence of anti-c and anti-E. All other major antibodies were ruled

Hsin-Yeh Hsieh, Diana L. Moroney, Deanne E. Naumann, D. Jane Hata, Nancy C. Vosnidou, Rovenna L. Kessinger, Nassir Shahab, Nasrollah Hakami, Daniel S. Smith

Immunohematology, Volume 18 , ISSUE 1, 19–22

Case report | 06-December-2020

Case report: serologic confirmation of paroxysmal cold hemoglobinuria

A child with a history of recent viral infection entered the hospital with severe anemia, hemoglobinuria, and suspected autoimmune disease. Serologic findings included a positive direct antiglobulin test and incompatible crossmatches. Extensive studies, including a Donath-Landsteiner test, confirmed paroxysmal cold hemoglobinuria. The child was transfused several times with washed red blood cells compatible by prewarm technique. Although hemolysis continued after each transfusion, he stabilized

Carol A. Putnam

Immunohematology, Volume 8 , ISSUE 1, 19–21

Article | 31-December-2020

An Example of Mild Hemolytic Disease of the Newborn Caused by Anti-Cob

A woman whose serum contained multiple alloantibodies delivered a full-term infant with mild hemolytic disease of the newborn. The direct antiglobulin test performed on the cord cells was positive with monospecific anti-IgG. An acid elution performed on the cord cells yielded anti-Cob. These findings were consistent with the presence of anti-Cob in the maternal serum. Neonatal clinical findings showed a mildly affected infant who demonstrated a moderate rise in total bilirubin and slight

Nancy B. Steffey, Mary A. Lieb

Immunohematology, Volume 3 , ISSUE 1, 9–10

Article | 10-November-2020

The impact of a gel system on routine work in a general hospital blood bank

The gel system has been reported to be more sensitive and specific than the conventional tube indirect antiglobulin test (IAT) for anti­body screening. However, a major concern about the gel system is its cost. A cost analysis study was therefore conducted at our hospi­tal. The gel system costs more than the conventional tube IAT per test; however, the total staff and reagent costs per year were about equal, because of staff savings. Workload and cost per patient requir­ing blood

A. Chan, H.F. Wong, C.H. Chui, L. Wong, G. Cheng

Immunohematology, Volume 12 , ISSUE 1, 30–32

Article | 17-November-2020

The first example of anti-Gya detected in Hong Kong

The incidence of anti-Gya is known to be extremely rare, and only a few examples have been reported in Japanese persons and in Caucasians. This case history reports the first example of anti-Gya detected in a Chinese person. The propositus is an 83-year-old male patient whose anti-Gya was revealed because of incompatibility of all units of blood tested by a saline indirect antiglobulin test (SIAT). Although the antibody had a titer of 512 by SIAT, 10 units of Gy(a+) red cells were transfused

King Hang Mak, Derek S. Ford, Gregory Cheng, Che Kit Lin, Cindy Yuen

Immunohematology, Volume 11 , ISSUE 1, 20–21

Article | 30-November-2020

Misidentification of anti-Vel due to inappropriate use of prewarming and adsorption techniques

Two units of red blood cells (RBCs) were ordered for a 44-year-old Caucasian woman with renal failure and cancer. Pretransfnsion testing performed at the regional reference laboratory had revealed the presence of an antibody reactive with all cells at the indirect antiglobulin test (IAT) but apparently nonreactive by a prewarmed IAT. The patient’s RBCs were direct antiglobulin test negative. Adsorption of the serum with rabbit erythrocyte stroma or with allogeneic RBCs at 4°C reduced

Jill Storry, Delores Mallory

Immunohematology, Volume 10 , ISSUE 3, 83–86

Article | 06-December-2020

Serologic investigation of fatal hemolytic anemia associated with a multiple drug history and Rh-like autoantibody

A patient who expired during an episode of gross intravascular hemolysis had a complex medical history, including renal disease, Coombs positive anemia of unclear etiology, recent transfusion, and cholecystectomy. Drug history included 21 different medications, including penicillin, acetaminophen, procainamide, furosemide, sulindac, and tolmetin, all of which have been associated with a positive direct antiglobulin test or drug-induced hemolytic anemia. The patient had a history of recent use

Nancy I. Maddox, Debra Futral, Floyd T. Boudreau

Immunohematology, Volume 8 , ISSUE 3, 70–76

Article | 06-December-2020

Failure to detect a prozoning anti-Fya in the serum of a chronically transfused patient

A 78-year old white male presented to our facility with vascular occlusion. He was Group A, D-positive, with a negative indirect antiglobulin test (antibody screen). Records at our facility, 4 years prior to this admission, indicated a history of anti-Fya, and Fy(a-) units were provided. A referring hospital had transfused the patient for "chronic anemia" (1-3 units weekly for 2 years), and he had received eight units (untested for Fya) immediately prior to his transfer to our

Jody C. Sizemore, Teresa D. Sammons, Jerry N. Clanton

Immunohematology, Volume 8 , ISSUE 2, 44–46

Article | 14-October-2020

Anti-Mta associated with three cases of hemolytic disease of the newborn

Carol C. Cheung, Daniel Challis, George Fisher, Susan J. Russell, Andrew Davis, Hayley Bruce, Julie Watt, Beng H. Chong

Immunohematology, Volume 18 , ISSUE 2, 37–39

Article | 26-October-2020

Precipitation of serum proteins by polyethylene glycol (PEG) in  pretransfusion testing

PEG was added. Rapid recognition of this phenomenon could prevent delay in the selection of blood for transfusion by substituting PEG-indirect antiglobulin test (IAT) with another technique such as low-ionic-strength solution and by increasing the number of washes prior to addition of the antiglobulin reagent.

Jack Hoffer, William P. Koslosky, Elizabeth S. Gloster, Therese M. Dimaio, Marion E. Reid

Immunohematology, Volume 15 , ISSUE 3, 105–107

Case Study | 10-November-2020

Characteristics of anti-Cob in vitro and in vivo: a case study

Serologic and biologic properties of an example of anti-Cob were investigated. The antibody was of the IgG class, and it bound small amounts of complement. It reacted optimally in the albumin-antiglobulin test with little or no enhancement of its reactivity in tests using enzymes. Additional experiments indicated that the Cob antigen is resistant to treatment with chemicals known to destroy other antigens. The antibody caused shortened survival of radiola­beled Co(b+) donor red cells in our

Johannes J.M.L. Hoffmann, Marijke A.M. Overbeeke

Immunohematology, Volume 12 , ISSUE 1, 11–13

Article | 14-December-2020

Indicators of clinically significant red cell antibodies produced by sensitized lymphocytes in liver transplant patients

It has been documented that transplanted livers can carry sensitized lymphocytes that subsequently produce red cell antibodies. We evaluated immunohematological variables in liver donors and recipients for indicators that might be predictive of serological red blood cell (RBC) destruction mediated by the passenger lymphocytes. Organ donor sera with antibody scores greater than ( > ) 60 correlated with a positive direct antiglobulin test (DAT) and need for increased RBC transfusion in liver

BeverIy E.W. Calhoun, M. Pothiawala, G. Musa, B. Baron

Immunohematology, Volume 7 , ISSUE 2, 37–39

Article | 14-October-2020

One thousand seventy antibodies detected only by a 2-stage papain test: wanted and unwanted positive reactions

Despite the wide use of the antibody detection test for unexpected antibodies, controversy still remains regarding the use of enzymetreated red blood cells. Over a 6-year period, 72,573 samples from 49,863 patients submitted for pretransfusion compatibility testing were examined for unexpected antibodies. The antibody detection tests included a low-ionic-strength solution (LISS) indirect antiglobulin test and a two-stage papain (2SP) test. One thousand and seventy of the 2267 (47%) antibodies

Carmen Martin-Vega, Dolores Castella, Joan Cid, Marta Panadés

Immunohematology, Volume 17 , ISSUE 4, 122–124

Case report | 30-November-2020

Case report: IgG1 Rh antibodies causing moderate hemolytic disease of the newborn

was group A, D positive, E negative, with a 3+ direct antiglobulin test. The eluate revealed anti-D and -hrB. Treatment of the hemolytic disease of the newborn included phototherapy, intravenous fluids, and transfusion of 60 mL of mother's deglycerolized red blood cells.

C. Faye Kugele, Cindy K. Oliver, Maria A. Carney, Jayne Hollander

Immunohematology, Volume 10 , ISSUE 4, 124–126

Article | 27-December-2020

The elucidation of a Kell-related autoantibody using ZZAP-treated red cells

A 61-year-old, nontransfused, Caucasian male was found to have a positive direct antiglobulin test (DAT) and an autoantibody in his serum prior to total hip replacement. Autoabsorption with the patient's ZZAP-treated red cells failed to absorb the autoantibody, giving a clue to its possible specificity, which was subsequently found to be Kell-system related. In addition, his red cells were found to have a slightly weakened expression of Kell system antigens. The patient was fit and healthy

Graham P. Rowe, Guiseppe G. Tozzo, Joyce Poole, Yew Wah Liew

Immunohematology, Volume 5 , ISSUE 3, 79–82

Article | 16-November-2020

A second example of anti-Esa, an antibody to a high-incidence Cromer antigen

A blood sample contained an antibody to a high-incidence antigen that reacted with all red blood cells (RBCs) tested by the indirect antiglobulin test (IAT). The antibody reacted with papain-, ficin-, and trypsin-treated RBCs, but not with α-chymotrypsin-treated RBCs. This pattern of reactivity suggested the possibility that the antibody was recognizing an antigen in the Cromer blood group system. Tests against RBCs deficient in decay-accelerating factor (which carries the Cromer antigens

Marion E. Reid, Roselyn Marfoe, Anita Mueller, Patricia A. Arndt, Laima Sausais, Peggy Spruell

Immunohematology, Volume 12 , ISSUE 3, 112–114

Case report | 06-December-2020

Case report: hemolytic anemia produced by tolmetin

Tolmetin, a nonsteroidal anti-inflammatory drug, was found to be the etiologic agent in a case of drug-induced hemolytic anemia. A 35-year-old female who had ingested tolmetin sporadically in the past took two doses that resulted in acute hemolysis. Two days after taking the second dose, she had a hemoglobin of 7.0 g/dL, increased serum lactate dehydrogenase, reticulocytosis, and indirect reacting hyperbilirubinemia. The direct antiglobulin test was weakly positive with anti-lgG and

LeeAnn McCall, Michael R. Owens

Immunohematology, Volume 8 , ISSUE 1, 17–18

Article | 30-November-2020

Autoimmune hemolysis following transfusion: a mimicking autoanti-D in a D- patient with alloanti-D

An 80-year-old group O, D- (rr) female with anti-C, -D, -E, and -Fya received four units of crossmatch-compatible red blood cells (RBCs). The direct antiglobulin test (DAT) was negative. Two weeks later, jaundice, dark urine, a 16% drop in hematocrit (Hct), a 20% reticulocyte count, and absent haptoglobin occurred. During the next month, her DAT was positive with anti-IgG and -C3d. Acid eluates, which repeatedly showed anti-D specificity, were nonreactive with enzyme-treated D- RBCs. Adsorption

Walter H. Dzik, Joyce Blank, Paula Lutz, Thomas G. Hirose, Christine Lomas-Francis, Marilyn Moulds

Immunohematology, Volume 10 , ISSUE 4, 117–119

Case report | 14-December-2020

Case report and review: alloimmunization, delayed hemolytic transfusion reaction, and clinically significant anti-Yta in a patient with ß-thalassemia/sickle cell anemia

were identified. The direct antiglobulin test was positive, and the eluate contained anti-c and -Jka. The patient’s hematocrit continued to decrease to 14 percent. Transfusions were withheld and the patient recovered uneventfully. Separate 51Cr red blood cell survival studies showed significantly shortened survival of both autologous and R1R1, Jk(a-), Yt(a+) erythrocytes. This case illustrates the complexity of transfusion management in hemoglobinopathy patients.

Christopher D. Hillyer, Jacquelynn M. Hall, Karen O. Tiegerman, Eugene M. Berkman

Immunohematology, Volume 7 , ISSUE 4, 102–106

Article | 14-December-2020

The sensitivity of antibody detection testing using pooled versus unpooled reagent red cells

. Little data, however, have been published to support this contention. In the present study, the data show a decreased sensitivity for antibody detection when pooled reagent RBCs are used. This reduced sensitivity could result in failure to detect some clinically significant RBC alloantibodies, which might result in the occurrence of overt hemolytic transfusion reactions, especially if an indirect antiglobulin test is not performed at the time blood is crossmatched.

Ira A. Shulman, Roland Nakayama, Cintia Calderon

Immunohematology, Volume 7 , ISSUE 1, 16–19

Case report | 29-December-2020

A case report: cold hemagglutinin disease in a pancreatic and renal transplant patient

thermal amplitude of 37°C. The patie  nt had a weakly positive direct antiglobulin test (DAT) with only complement detectable on the red cells. The patient recovered spontaneously. The etiology of the CHD is unclear. The use of cyclosporin-A, OKT3 monoclonal antibody, and anti-thymocyte globulin (ATG) to treat acute rejection could have played a part.

Catherine Y. Beiting, Kathleen S. Larimore

Immunohematology, Volume 4 , ISSUE 4, 85–87

Article | 14-October-2020

Evaluation of a new solid-phase immunoassay for alloantibody detection using bromelin-treated and untreated red blood cells

detected with enzyme-treated intact RBCs and untreated RBCs by M-MPHA. The slight increase in reactivity using M-MPHA was not seen using dried RBC stroma (M-MPHA-Dry). All donor-derived IgG alloantibodies, which were detected by either a conventional tube enzyme test or an indirect antiglobulin test, were detected by M-MPHA without using enzyme-treated RBCs. Both M-MPHA and M-MPHA-Dry can be used for antibody detection without using enzyme-treated RBCs and are also useful for antibody identification.

Toyohiro Tamai, Toshio Mazda

Immunohematology, Volume 17 , ISSUE 1, 17–21

Case report | 16-November-2020

Case report: rapid onset of severe anemia in childhood. A systematic approach to laboratory diagnosis

Evaluation of a patient prompted the systematic review of causes of abrupt onset anemia in children and the development of a protocol for the immunohematologic workup of such cases. The patient, a 3- year-old boy, presented to our institution two weeks following a viral illness with gross hematuria, jaundice, and severe anemia (hematocrit 14.5%). He was found to have a positive direct antiglobulin test (DAT) due to primarily complement sensitization (2+) with smaller amounts of IgG (weak). A

Linda A. Chambers

Immunohematology, Volume 12 , ISSUE 3, 108–111

Article | 06-December-2020

Reactive lysis-a phenomenon of delayed hemolytic transfusion reactions

posttransfusion, a mixed-field anti-IgG direct antiglobulin test (DAT) indicated that there were two cell populations present. The DAT remained positive with anti-C3b, -C3d throughout the course of the reaction. Because of the substantially lower Hb and the severity of symptoms, immunologic clearance of the antigen-positive donor RBCs was expected. However, we demonstrated their persistence. Our data corroborates the observation that complement activation occurring during DHTRs can result in the hemolysis of

Deborah L. Greene, Sanobar Khan

Immunohematology, Volume 9 , ISSUE 3, 74–77

Article | 14-December-2020

Antibody detection using pooled sera and a solid phase system

The purpose of the study was to evaluate the feasibility of substituting the Immucor Capture™-R solid phase (SP) antibody detection system for our routine donor antibody screen. Our routine procedure (RP) used a 12-drop pool of six donor sera and one drop of pooled reagent red cells, with 37°C incubation and indirect antiglobulin test readings. The SP system was used according to the manufacturer’s directions except that one drop of the pooled sera (rather than an individual

Malcolm L. Beck, Jill T. Hardman, Alicia M. Briseño

Immunohematology, Volume 7 , ISSUE 3, 73–75

Case report | 20-December-2020

A case report: IgG autoanti-N as a cause of severe autoimmune hemolytic anemia

A 21-year-old white woman was referred for evaluation of hemolytic anemia after a 9-day history of marked hemoglobinuria, jaundice, and weakness. The patient’s hematocrit was 18%, despite at least eight transfusions over the previous week, and the reticulocyte count was <1%. Serologic evaluation revealed a weakly positive direct antiglobulin test with antic-C3 only. The serum contained cold-and warm-reacting anti-N. Dithiothreitol had no effect on either the cold- or the warm-reacting

Martha Rae Combs, Marilyn J. Telen, Sharon E. Hall, Wendell F. Rosse

Immunohematology, Volume 6 , ISSUE 4, 83–86

Review | 01-December-2019

Cold acid elution (ELU Kit II)

Elution is a procedure for recovery of antibody attached to intact, immunoglobulin-coated red blood cells (RBCs) by disrupting the antigen–antibody bonds. The recovered antibody is collected in an inert diluent and is referred to as an eluate. Testing of an eluate may be desired to identify antibody(ies) coating the RBCs of patients with a positive direct antiglobulin test. Many types of elution procedures have been developed and described; however, an acid elution is suitable for

Monica Hinrichs, Monica A. Keith

Immunohematology, Volume 30 , ISSUE 3, 113–116

Article | 18-October-2020

Human anti-Dia monoclonal antibodies for mass screening

The use of monoclonal antibodies (mabs) to blood group antigens is constantly increasing for routine typing. Two heterohybridoma cell lines, HMR15 and HMR22, were established by Epstein-Barr virus transformation of peripheral blood lymphocytes from a blood donor with anti-Dia. HMR15 mab directly agglutinated Di(a+) red cells, and HMR22 mab agglutinated Di(a+) red cells exclusively by the indirect antiglobulin test. Reactivities of both HMR15 and HMR22 mabs were specific for Dia and had good

Toru Miyazaki, Shinichiro Sato, Toshiaki Kato, Hisami Ikeda

Immunohematology, Volume 16 , ISSUE 2, 78–81

Article | 10-November-2020

The second example of Lu:–7 phenotype: serology and immunochemical studies

We describe the second example of red blood cells (RBCs) with the Lu:–7 phenotype in a 37-year-old Latino female (SA). Her RBCs were nonreactive with anti-Lu7 (Mrs. GA) but were reactive with all other antibodies to high-prevalence antigens tested, including those in the Lutheran blood group system. No Lu:–7 RBCs were available for testing. SA’s serum was nonreactive by the indirect antiglobulin test against (1) recessive and dominant Lu(a–b–) RBCs and (2) trypsin

Marion E. Reid, Jack L. Hoffer, Ragnhild Øyen, Edith Alicea-Tossas, Manijeh Sadjadi, Gladys M. Messina

Immunohematology, Volume 12 , ISSUE 2, 66–68

Article | 30-November-2020

An example of anti-LWa in a 10-month-old infant

Blood samples from a 10-month-old male infant requiring transfusion were found to contain an allomtibody reacting at 37°C in saline, by indirect antiglobulin test (IAT), and with a manual polybrene technique. Preliminary results suggested anti-D and another weaker reacting antibody, but the patient had been previously transfused with only D- blood. His serum reacted more weakly by IAT against red cells treated with 0.2M dithiothreitol (DTT), and one D+, LW(a-) sample was nonreactive. The

Alan Devenish

Immunohematology, Volume 10 , ISSUE 4, 127–129

Article | 06-December-2020

Antibody detection errors due to acidic or unbuffered saline

Isotonic saline solutions, buffered with potassium phosphate or sodium phosphate salts, were evaluated in parallel with unbuffered saline to determine if they improved antibody detection by solid phase red cell adherence or hemagglutination methods. Saline buffered to a pH of 7.0 to 7.5, when used to suspend red cells or to wash sensitized red cells in preparation for the antiglobulin test, produced the best positive solid phase and hemagglutination results. The pH range of commercially

Susan Rolih, Ron Thomas, Fern Fisher, Joanne Talbot

Immunohematology, Volume 9 , ISSUE 1, 15–18

Case report | 09-October-2019

Autoanti-C in a patient with primary sclerosing cholangitis and autoimmune hemolytic anemia: a rare presentation

Primary sclerosing cholangitis (PSC) is rarely associated with autoimmune hemolytic anemia (AIHA), and the presence of  specific autoantibodies has not been reported previously. We present a unique case report of PSC associated with AIHA implicating autoanti-C. A 17-year-old girl was admitted to our hospital with PSC along with AIHA. Her blood sample demonstrated a positive direct antiglobulin test and a positive autocontrol in the antihuman globulin phase, confirming the patient had warm

Meenu Bajpai, Ashish Maheshwari, Shruti Gupta, Chhagan Bihari

Immunohematology, Volume 32 , ISSUE 3, 104–107

Case report

Weak D type 67 in four related Canadian blood donors

Correct donor D typing is critical to prevent recipient alloimmunization. No method can detect all variants, and the immunogenicity of many variants is unknown. Routine ABO and D serologic typings are performed in our laboratory by automated microplate testing. Until 2011, routine confirmation of D– status of first-time donors was performed by the manual tube indirect antiglobulin test (IAT); this was replaced by automated solidphase testing including weak D testing by IAT. Selected

Philip Berardi, Emma Bessette, Michiko Ng, Nancy Angus, Debra Lane, Lise Gariepy, Katerina Pavenski, Gorka Ochoa-Garay, Jacqueline Cote, Mindy Goldman

Immunohematology, Volume 31 , ISSUE 4, 159–162

Case report

Blocked D phenomenon and relevance of maternal  serologic testing

A blood requisition for double-volume exchange transfusion was received for a 2-day-old male child born to a 29-yearold multiparous female (P2002) referred to our institute having neonatal jaundice with encephalopathy; no maternal sample was received. The neonatal blood sample was typed as group A, D–, and the direct antiglobulin test (DAT) was strongly positive (4+) using the gel method. Mono-specific DAT showed the presence of IgG antibodies on neonatal red blood cells (RBCs). Acid

Ashish Jain, Vijay Kumawat, Neelam Marwaha

Immunohematology, Volume 31 , ISSUE 3, 116–118

Case report | 01-December-2019

A case of masquerading alloantibodies:  the value of a multitechnique approach

, D+ blood type showing strong reactivity with all cells tested  in the forward and reverse ABO, in the D testing as well as in a three-cell antibody screen. The initial assumption was that the plasma contained a cold autoantibody. Subsequent testing, including the use of gel column technology, ficin-treated cells, and antisera for phenotyping, showed the apparent cold autoantibody to be a red herring. Additional tube testing at immediate spin, 37°C, and indirect antiglobulin test (IAT

Paula M.S. Wennersten, Laurie J. Sutor

Immunohematology, Volume 30 , ISSUE 3, 117–120

Article | 14-October-2020

Antibody screening in 37°C saline. Is it safe to omit it using the indirect antiglobulin (gel) test?

Pretransfusion tests must detect antibodies that can shorten the life of red blood cells (RBCs). Some studies have demonstrated the existence of clinically significant antibodies detected at 37°C in saline that are not detected by the indirect antiglobulin test (IAT) when the conventional tube test is used. Our aim was to determine whether these antibodies, detected with a 37°C saline tube test, are also detected when a sensitive column gel agglutination method is used. The 2373

José A. Duran, Manuel Figueiredo

Immunohematology, Volume 18 , ISSUE 1, 13–15

Article | 26-October-2020

DNA from urine sediment or buccal cells can be used for blood group molecular genotyping

Accurate blood group antigen typing of red blood cells with a positive direct antiglobulin test or from a recently transfused patient has been a long-standing problem. To overcome this problem, we evaluated the feasibility of using somatic cells as a source of DNA for molecular genotyping. Two sources of cells that could be obtained by noninvasive procedures were chosen for analysis: urine samples, which were already available in the clinical laboratory, and buccal epithelial cells collected

Marion E. Reid, Maria J. Rios, Kevin L. Cash, Annie M. Strupp, Joan M. Uehlinger

Immunohematology, Volume 15 , ISSUE 2, 61–65

Article | 10-November-2020

Anti-Holley detected in a primary immune response

. The antibody was reactive at room temperature, 37°C, and in the antiglobulin phase. IgG and IgM components of anti-Hy were demonstrated in the maternal serum, documenting a primary immune response. This resulted in serologic findings not previously described for anti-Hy. A direct antiglobulin test on the newborn red cells was negative and there was no clinical evidence of hemolytic disease of the newborn (HDN). A monocyte monolayer assay performed with maternal serum yielded negative results

Vicki J. Barrett, M. Margaret O’Brien, John J Moulds, Peggy Spruell, Valerie Jackson, James R. Stubbs

Immunohematology, Volume 12 , ISSUE 2, 62–65

Article | 17-November-2020

A comparison oftwo solid phase systems for antibody detection

Two solid phase methods of antibody detection, Capture-R (CR) and Capture-R Ready-Screen (RS), were compared to determine their acceptability for use in prenatal antibody screening. Ninety-six serum samples, screened using a saline antiglobulin test, were coded and tested by CR and RS at two laboratory sites using a blinded study design. Thirty of the samples were free of antibody, and 66 samples contained antibody. Parallel testing was also performed in both laboratories on 648 prenatal

Gwen M. Haslam, Margaret Persaud, Yvette Fournier, Joanne Sajur, Janice Aulph, Nancy Heddle

Immunohematology, Volume 11 , ISSUE 1, 8–10

Article | 30-November-2020

Hemolytic transfusion reactions due to anti-e+f detectable only by nonstandard serologic techniques

. The patient’s RBCs typed as A, D+, probable Rh phenotype (cDE/cDE). The direct antiglobulin test was negative, and serum samples following the second transfusion were red/brown in color. Serologic investigations were inconclusive on all samples taken until February 13 (after the fourth transfusion). At this time, a weak anti-e reacting by manual polybrene technique and an anti-e+f reacting by two-stage papain technique were detected. The serum also contained potent HLA antibodies. The

Alan Devenish, Lesley A. Kay

Immunohematology, Volume 10 , ISSUE 4, 120–123

Article | 31-December-2020

Cold Autoimmune Hemolytic Anemia With Auto-Anti-AI Specificity: 51Chromium Studies

A 65-year-old woman was found to have severe autoimmune hemolytic anemia. The patient was group A1, RhoD positive. The direct antiglobulin test was strongly positive with anti-C3 and negative with anti-IgG. The serum contained two distinct IgM antibodies, auto-anti-I and auto-anti-AI. Both were reactive at 22°C. However, the anti-AI also was reactive in saline and in albumin at 37°C. An eluate revealed anti-AI and a weak anti-I. Sequential 51Chromium survival studies were done with

May H. McGinniss, Richard J. Davey, Richard A. Binder, Arthur N. Kales

Immunohematology, Volume 3 , ISSUE 2, 20–22

Case report | 09-October-2019

Hemolytic transfusion reaction attributable to anti-Dia  

In situations when a patient's antibody detection test is negative, many institutions have moved from an indirect antiglobulin test (IAT) crossmatch to an electronic crossmatch system. Here we report a case of an acute hemolytic transfusion reaction attributable to anti-Dia in a patient with a negative antibody detection test. A 22-year-old female patient with a diagnosis of β thalassemia and sickle cell anemia commenced a routine exchange transfusion of 5 units of red blood cells

Arthur J. Joyce, Kelli M Quantock, Ray Banh, Yew-Wah Liew

Immunohematology, Volume 33 , ISSUE 1, 6–8

Case report | 14-October-2020

Discrepancies in Rh(D) typing of sensitized red blood cells using monoclonal/polyclonal anti-D reagents: case report and review

antiglobulin test and, in some samples, spontaneous agglutination in saline. One intent of the surveys was to monitor the accuracy of the reported Rh(D) type. On an initial survey, 19 of 115 (16.5%) participants reported the RBCs as D+. Of these laboratories, 63.2 percent (12/19) had used a monoclonal/ polyclonal blend anti-D reagent. On a subsequent survey, after educational material had been distributed, only five of 113 (4.4%) participants reported the Rh type as D+. Two of these five laboratories had

Beverly J. Padget, Judith L. Hannon

Immunohematology, Volume 17 , ISSUE 1, 10–13

Article | 18-October-2020

Comparison of tube and gel techniques for antibody identification

There are several methods for antibody detection and each technique has advantages and limitations. We compared the performance of the tube (polyethylene glycol–indirect antiglobulin test [PEG-IAT]) and gel test technique for antibody identification. From January to May 1999, we performed antibody screening tests by gel and tube techniques on 10,123 random blood samples submitted to our reference laboratory. Six hundred and twentyeight (6.2%) reactive samples were tested for antibody

Marcia Cristina Zago Novaretti, Eduardo Jens Silveira, Edio da Costa Filho, Pedro Enrique Dorlhiac- Llacer, Dalton de Alencar Fischer Chamone

Immunohematology, Volume 16 , ISSUE 4, 138–141

Article | 18-October-2020

A successful delivery of a baby from a D––/ D–– mother with strong anti-Hr0

, fetal cells were direct antiglobulin test positive and the blood type was group O, D+ (CDe). We performed plasma exchanges in the mother; however, the titer of antibody rebounded to its initial level after the third plasma exchange. At 26 weeks gestation, cord blood Hb decreased to 7.1 g/dL and three intrauterine transfusions were performed using the mother’s washed red blood cells (RBCs). At 34 weeks gestation, a live baby was delivered by cesarean section. The infant was hydropic with

Dong Hee Whang, Hee Chung Kim, Mina Hur, Jung Hwan Choi, Joong Shin Park, Kyou Sup Han

Immunohematology, Volume 16 , ISSUE 3, 112–114

Article | 26-October-2020

Anti-Lu9: the finding of the second example after 25 years

The first and only reported exanipie of anti-Lu9 (an antibody directed at a low-incidence antigen in the Lutheran blood group system and allelic to the high-incidence antigen Lu6) was described in 1973 in the serum of a white female, Mrs. Mull. Her serum also contained anti-Lul1 (-Lua), and subsequently, an anti-HLA-B7 (-Bga) was identified. We report the second example of anti-Lu9 in a white male (GR), found 25 years later. The GR serum was reactive in the indirect antiglobulin test with Lu

Kayla D. Champagne, Marilyn Moulds, Jo Schmidt

Immunohematology, Volume 15 , ISSUE 3, 113–116

Article | 03-November-2020

Paroxysmal cold hemoglobinuria and the elusive DonathLandsteiner antibody

Four patients with paroxysmal cold hemoglobinuria (PCH) illustrate some of the difficulties in making the diagnosis. A 46-year-old male presented with anemia, a weakly positive direct antiglobulin test (DAT) with anti-IgG, a haptoglobin < 0.1 g/L, and a cold autoagglutinin showing anti-P specificity. A 9-year-old female had a 4-day coryzal illness, a 20 g/L fall in hemoglobin over 24 hours, and a haptoglobin < 0.1 g/L; the DAT was positive with anti-C3d. A 3-year-old female was referred

R.J. Sokol, D.J. Booker, R. Stamps

Immunohematology, Volume 14 , ISSUE 3, 109–112

Article | 09-November-2020

Delayed hemolytic transfusion reaction and paroxysmal cold hemoglobinuria: an unusual association

An 80-year-old female presented with melena and anemia due to bleeding from a benign gastric ulcer. Her blood group was O, D+. The serum contained anti-B and a weak anti-A (titer 2 at 18°C). She was inadvertently transfused with approximately 3.5 units of group A red blood cells with no initial ill effects. One week later, the antiA titer increased to 8 and the direct antiglobulin test (DAT) was weakly positive (IgG and C3d). The next day, intravascular hemolysis became evident. The DAT was

M.A. Wodzinski, R.C. Collin, D.J. Booker, R. Stamps, J.D. Bellamy, R.J. Sokol

Immunohematology, Volume 13 , ISSUE 2, 54–57

Article | 17-November-2020

Analysis ofthe routine use of polyethylene glycol (PEG) as an enhancement medium

This study compared the performance of polyethylene glycol (PEG) and low-ionic saline solutions (LISS) as enhancement media for routine use in a large transfusion service. A PEG additive solution (PEG plus LISS) was compared to a LISS additive (LISS plus polymers) and to an albumin-indirect antiglobulin test (A-IAT). Fifty serum samples containing clinically significant alloantibodies and fifty samples without alloantibodies were tested. Following an acute hemolytic transfusion reaction (HTR

Vicki J. Barrett, James R. Stubbs, Karen Stuardi, Angela Hollis, Leslie Clear

Immunohematology, Volume 11 , ISSUE 1, 11–13

Article | 30-November-2020

Successful transfusion in the presence of anti-K4 (anti-Kpb)

Type, Bristol, UK, maintained by the World Health Organization. Four units of group O, K:4, E-, S- RBCs were transfudes in the next 24 hours while the K:-4 blood was in transit. No immediate signs of a transfusion reaction were noted. Blood pressure, temperature, bilirubin, and urinary output were normal, and an increase in hemoglobin was observed. A positive direct antiglobulin test was evident 3 days posttransfusion of the incompatible units and was still present 8 days later. Anti-A and anti-K4

Julie M Watt, Peter N. Moffatt, Suzanne Y. Chatfield, Wendy A. Grimm, Jennifer A. Bryant

Immunohematology, Volume 10 , ISSUE 3, 87–89

Article | 06-December-2020

Anti-Uz found in mother's serum and child's eluate

A saline-reactive antibody, anti-Uz, that reacted stronger with S+ than with S- red blood cells (RBCs) and failed to react with U- or ficin-treated RBCs has been previously reported. We describe an antibody of similar specificity in the postpartum serum of an untransfused woman and the eluate from her fourth child's cord RBCs. The mother's RBCs typed S-s+U+, He+(weak), and appeared to have normal glycophorin A and B content, as determined by immunoblotting. The direct antiglobulin test

Sandra M. Read, Mary M. Taylor, Marion E. Reid, Mark A. Popovsky

Immunohematology, Volume 9 , ISSUE 2, 47–49

Article | 14-December-2020

Primary immune response to blood group antigens in burned children

positive direct antiglobulin test (DAT), or both. None of the 11 patients included in the study had been previously tranfused or pregnant. The average number of units transfused prior to antibody identification was 19. The average time elapsed between the first transfusion and antibody identification was 3.6 weeks. Anti-K and anti-E were the most frequently identified. Three patients had a decrease in hemoglobin (average 1.5 g/dL) and hematocrit at the time that a positive DAT was detected. Such

Nancy E. Bacon, Ethel D. Patten, Janet L. Vincent

Immunohematology, Volume 7 , ISSUE 1, 8–11

Article | 14-October-2020

MIMA-9, a valuable antibody for screening for rare donors

and thereby conserve rare antisera. Inclusion of anti-mouse IgG in gel cards allowed for rapid screening. MIMA-9 is also a useful reagent to type RBCs with a positive direct antiglobulin test. This antibody is available to donor screening laboratories at no cost for this specific use.

Edith Tossas, Ragnhild Øyen, Gregory R. Halverson, Harry Malyska, Marion E. Reid

Immunohematology, Volume 18 , ISSUE 2, 43–45

Article | 14-October-2020

Tube and column agglutination technology for autocontrol testing

agglutination tests. The tube method was carried out using low-ionic-strength solution (LISS). The direct antiglobulin test (DAT) was performed using the tube method, and further investigated with elution studies if warranted. Seventy-nine patient’s samples (7.74%) had a positive autocontrol: the gel test, 72 (91.13%); ReACT, 21 (26.58%); and the tube method, 27 (34.18%). Of the 79 positive autocontrols, 44 samples had a negative DAT. Of the samples with positive DAT results, only one possessed a

J.E. Courtney, J.L. Vincent, A.J. Indrikovs

Immunohematology, Volume 17 , ISSUE 2, 50–52

Article | 09-November-2020

Direct Coombs test-negative autoimmune hemolytic anemia and low-affinity IgG class antibodies

Autoimmune hemolytic anemia, in which the direct antiglobulin test (DAT) is negative or weakly positive, may be due to low-affinity autoantibodies. We describe two such cases. An 8-year-old male presented with weight loss, jaundice, a hemoglobin of 33 g/L, reticulocytes of 306 x 109/L, and haptoglobin of < 0.1 g/L. The DAT was negative. After washing the red blood cells (RBCs) with saline at 4°C, the DAT was positive for IgG and an eluate contained an IgG3 autoantibody, thus confirming a

R.J. Sokol, D.J. Booker, R. Stamps, S. Jalihal, B. Paul

Immunohematology, Volume 13 , ISSUE 4, 115–118

Article | 17-February-2021

Prevalence of DEL phenotype in D– blood donors in India

samples negative for D by the indirect antiglobulin test (IAT) were found to be positive by adsorption-elution.3 Until recently, DEL was primarily described in Asians, since a significant proportion of D– Asian individuals are in fact DEL+.5–7 In Caucasians, DEL is very rare, although recent molecular studies show that the prevalence of DEL in donors typed D– in routine testing is greater than initially thought.8,9 Molecular studies have shown that a heterogeneous array of variant RHD alleles can

R. Chaudhary, S. Verma, A. Verma

Immunohematology, Volume 36 , ISSUE 4, 133–136

Case report | 14-October-2020

Moderate hemolytic disease of the newborn (HDN) due to anti-Rh17 produced by a black female with an e variant phenotype

antiglobulin test was 2+ with anti-IgG. Total bilirubin rose to 14.2 mg/dL within 48 hours. Indirect bilirubin peaked at 14.7 mg/dL. The bilirubin responded to triple phototherapy. The infant was discharged on day 6. Potential for infant morbidity due to anti-Rh17- mediated HDN and the importance of specifying risks to women with this antibody if they contemplate pregnancy are discussed.

Marla C. Brumit, Gary E. Carnahan, James R. Stubbs, Jill R. Storry, Marion E. Reid

Immunohematology, Volume 18 , ISSUE 2, 40–42

Article | 09-November-2020

The use of polyethylene glycol (PEG) to enhance the adsorption of autoantibodies

RBCs, and PEG for 15 minutes at 37°C. The PEG/serum mixture was harvested and used for testing. Six drops of the PEG/serum mixture were tested against reagent RBCs for 15 minutes at 37°C. An antiglobulin test was then performed using anti-IgG. The PEG adsorption technique took a total of 10 hours to completely eliminate autoantibody reactivity in all 19 samples. The reference method required a total of 59.5 hours to adsorb the autoantibodies in all 19 samples. Two weak alloantibody

Christina L. Barron, Mary Beth Brown

Immunohematology, Volume 13 , ISSUE 4, 119–122

Article | 10-November-2020

Do monocyte ADCC assays accurately predict the severity of hemolytic disease of the newborn caused by antibodies to high-frequency antigens?

Monocyte ADCC assays are helpful indicators of the severity of hemolytic disease of the newborn (HDN) due to anti-D. It would be particularly useful if the assays also accurately predicted the ability of antibodies to high-frequency antigens (HFA) to cause HDN. To investigate this possibility, 14 antenatal sera containing antibodies to HFA were tested and the results correlated with the severity of HDN. Antibody titers were determined using an indirect antiglobulin test (IAT). Eight sera 

Stephen F. Garner, Alan Devenish

Immunohematology, Volume 12 , ISSUE 1, 20–26

Case report | 09-October-2019

Development of red blood cell autoantibodies following treatment with checkpoint inhibitors: a new class of anti-neoplastic, immunotherapeutic agents associated with immune dysregulation

chart review, including serologic, hematology, and chemistry laboratory results, of two patients who developed red blood cell (RBC) autoantibodies during treatment with a checkpoint inhibitor. Serologic testing of blood samples from these patients during induction therapy with ipilimumab and nivolumab, respectively, showed their RBCs to be positive by the direct antiglobulin test (IgG+, C3+) and their plasma to contain panreactive RBC autoantibodies. Neither patient had evidence of hemolysis. Both

Laura L.W. Cooling, John Sherbeck, Jonathon C. Mowers, Sheri L. Hugan

Immunohematology, Volume 33 , ISSUE 1, 15–21

Case report | 09-October-2019

Two cases of the variant RHD*DAU5 allele associated with maternal alloanti-D  

allele. Cord blood testing on both infants revealed positive direct antiglobulin test (DAT) results with anti-D eluted from the red blood cells (RBCs) of one of the infants. Despite the positive DAT, neither infant experienced anemia or hyperbilirubinemia. We document two cases of pregnant women whose RBCs expressed a partial D variant and were classified as D+ on the basis of standard serologic testing, resulting in subsequent failure to provide RhIG prophylaxis. Both cases were associated with

Jennifer A. Duncan, Susan Nahirniak, Rodrigo Onell, Gwen Clarke

Immunohematology, Volume 33 , ISSUE 2, 60–63

Report | 01-December-2019

Identifying D-positive donors using a second automated testing platform

estimated the number of cases of alloimmunization in women younger than 50 years likely to be prevented by the addition of Galileo testing. From May 2011 to May 2012, 910,220 donor samples were tested; 15,441 were first-time donors with concordant D– results. Five donors tested D– on the PK7300 and weak D+ on the Galileo; one was found to be a false positive on further testing. On manual testing, the other four donors had positive indirect antiglobulin test results with one to three of the

Mindy Goldman, Ilona Resz, Jacqueline Cote, Gorka Ochoa, Nancy Angus

Immunohematology, Volume 29 , ISSUE 3, 97–100

Article | 03-November-2020

Use of LOR-15C9 monoclonal anti-D to differentiate erythrocytes with the partial DVI antigen from those with other partial D antigens or weak D antigens

Historically, red blood cells (RBCs) with partial D antigens have been defined serologically by their pattern of reactivity with polyclonal and monoclonal anti-D. Although numerous variants have been described in tests with well-characterized monoclonal anti-D, definition remains difficult to ascertain serologically. RBCs of known partial D type were tested with LOR-15C9 (a monoclonal anti-D) and commercial anti-D by the tube indirect antiglobulin test (IAT), by micro typing system IgG gel

Marion E. Reid, Gregory R. Halverson, Francis Roubinet, P.A. Apoil, Antoine Blancher

Immunohematology, Volume 14 , ISSUE 3, 89–93

Article | 03-November-2020

Autoimmune hemolytic anemia caused by warm-reacting IgM-class antibodies

confirmed as IgM by their ability to rebind to normal red blood cells (RBCs) after elution; the absence of small increases in RBC-bound IgG and IgA was shown by a sensitive enzyme-linked antiglobulin test. Patient 1 was a 64-year-old female with non-Hodgkin’s lymphoma, with a hemoglobin of 50 g/L and haptoglobin of < 0.1 g/L. Direct antiglobulin tests were positive for IgM, C3d, and C3c; only IgM was present in an eluate. The serum contained a weak autoantibody at 37°C and tests for

R.J. Sokol, D.J. Booker, R. Stamps, S. Sobolewski, A.P. Haynes

Immunohematology, Volume 14 , ISSUE 2, 53–58

Article | 22-November-2020

Cefotetan-induced immune hemolytic anemia due to the drug-adsorption mechanism

epinephrine, and she was transfused with an additional unit of RBCs. Her Hct rose to 34.1% prior to her discharge on day 11. Further investigation revealed that the patient's serum and an eluate contained an antibody that reacted with cefotetan- and cephalothin-coated RBCs by the indirect antiglobulin test. In a monocyte monolayer assay, monocytes readily phagocytized cefotetan- and cephalothin-coated reagent RBCs but not uncoated reagent RBCs. The patient's serum did not react by the so-called

Robert J. Eckrich, Susan Fox, Delores Mallory

Immunohematology, Volume 10 , ISSUE 2, 51–54

Case report | 16-March-2020

Autoantibody formation after alloimmunization inducing bystander immune hemolysis

pregnancies 40 years before, but no history of RBC transfusion, and her antibody screen was negative. On the tenth day after transfusion her hemoglobin dropped, and alloanti-c was identified in her serum and eluate. At this time she received another two units of compatible blood according to her phenotype (group O, R1R1, K:-1). After 48 hours, she developed joint pain, pyrexia, and hemoglobinuria, and her Hb dropped from 9.2 g/dL to 5.3 g/ dL. The direct antiglobulin test was positive, an IgG autoantibody

Mariza Mota, C. Bley, M.G. Aravechia, N. Hamerschlak, A. Sakashita, J.M. Kutner, L. Castilho

Immunohematology, Volume 25 , ISSUE 1, 9–12

Case report | 01-December-2019

Molecular analysis of patients with weak D and serologic analysis of those with anti-D (excluding type 1 and type 2)

included autologous controls, direct antiglobulin test, elution, and titration of anti-D before and after adsorption of serum onto autologous RBCs. From molecular analyses, 459 individuals exhibited a weak D type. We described seven novel RHD variant alleles. The most frequent types of weak D were type 1 (30.1%), type 2 (23.7%), type 4.0 (10.2%), type 4.2.2 (20.3%), type 11 (3.9%), and type 15 (3.7%). Anti-D was identified in the sera of 9 of 47 individuals with weak D type 4.0, in 14 of 93 with weak D

Bach-Nga Pham, Michèle Roussel, Dominique Gien, Maryline Ripaux, Carine Auxerre, Pierre-Yves Le Pennec, Christine Andre-Botte

Immunohematology, Volume 29 , ISSUE 2, 55–62

Article | 03-November-2020

Improved detection of weak, clinically significant antibodies by supplementation of polyethylene glycol with a low-ionic solution

A comparative study of 164 serum samples was carried out to determine the specificity and sensitivity of the indirect antiglobulin test (IAGT) in three different formulations: physiologic saline, low-ionic solution (RAM), and RAM supplemented with polyethylene glycol (PEG). Serum samples containing mostly weak antibodies (anti-D, -C, -E, -c, -Jka, -Fya, -K, -S, -Lea, -Lua, -M, -Cob, -P1, -I, and -Kna) were used in a 10-minute IAGT in which PEG-IAGTs were compared with salineIAGTs and RAM-IAGTs

Kim Swee Low, Yew-Wah Liew, Peter M. Bradley

Immunohematology, Volume 14 , ISSUE 2, 68–71

Article | 22-January-2021

Routine indirect antiglobulin testing of blood donors—a further step toward blood safety: an experience from a tertiary care center in northern India

transfusions, since the presence of unexpected antibodies in the donor plasma can lead to adverse transfusion reactions, particularly in cases in which a large amount of plasma is transfused (as in massive transfusions) and in pediatric patients.3 In this scientific article, we emphasize the importance of establishing a policy for antibody screening of all blood donors as a step to further improve blood safety. The policy of routine inclusion of the indirect antiglobulin test (IAT) was adopted in our

S. Malhotra, G. Negi, D. Kaur, S.K. Meinia, A.K. Tiwari, S. Mitra

Immunohematology, Volume 36 , ISSUE 3, 93–98

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